Abstract An improved and efficient protocol was developed based on the
TaKaRa RNAiso Plus Kit (Code: D9108A) for isolating good-quality total
RNA from the optic stalk of mud crab, Scylla paramamosain . The
protocol was based on the Trizol method with modifications. The
carapace overlapping the optic stalk was retained with RNA in regular
protocol. In order to remove the abundant deposition correlative with
the carapace which makes the isolation of RNA particularly difficult,
5M potassium acetate solution (pH = 6.0) was added before the
precipitation of RNA, and the temperature of RNA deposition was also
decreased to -70\ubaC to ensure the stabilization of RNA.
Good-quality total RNA from the optic stalk of S. paramamosain could be
easily isolated with this modified protocol and three conventional
methods were also employed to confirm the quality of RNA. This improved
method would be helpful in facilitating molecular research of crabs
involving RNA from the optic stalk